Soft x-ray microscopy shows enormous promise as a technique for imaging cellular structures at resolutions well beyond what can be achieved in optical microscopes, and with much simpler sample preparation than is required for electron microscopy. In addition, the lower radiation dose required (compared to electron microscopy) allows tomographic investigation of subcellular structures in three dimensions. Existing synchrotron based microscopes have shown the potential of the soft x-ray microscope as a research tool, but they have the disadvantage of being tied to a massive light source at only a few national laboratories. A suitably bright, compact, low cost light source is the enabling innovation needed to realize a commercial biological soft x-ray microscope. In a Phase I NIH SBIR program (2R44RR022488-02) Energetiq demonstrated a unique, compact light source that generates the required x-ray wavelength and intensity for imaging. In the following Phase II NIH SBIR program (5R44RR022488-03) Energetiq constructed a "proof-of-principal" microscope. With three months left to go in the Phase II program, nearly all major program goals have been met, and we are on track to completing the rest of the program goals on schedule and within budget. In the proposed Phase II Renewal program, Energetiq will focus on significantly shortening exposure time through increasing the source brightness and will work with an existing manufacturer (Xradia, Inc.) of x-ray microscopes to integrate the Energetiq source with their microscope, thus accelerating the point at which a commercial instrument will be available in the market. The ultimate goal will be a new commercial tool for researchers to use in studying subcellular structures. PUBLIC HEALTH RELEVANCE: Biologists have a need to image whole biological cells at high resolution and in their hydrated state. Currently available commercial, integrated microscopes satisfy one of these requirements but not both. Soft x-ray microscopy in the so-called 'water window'allows resolution to 20nm for fully hydrated cell samples, and can produce 2-D and 3-D tomographic images. The ability to resolve subcellular structures in hydrated cells, and to create those images in minutes rather than days, could lead to more rapid advances in drug discovery, disease diagnosis, disease treatment and in the understanding of fundamental cellular processes. The proposed program will enable the widespread adoption of this technique by researchers.